Difference between revisions of "Part:BBa K1365006:Design"

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===Design Notes===
 
===Design Notes===
−
Used primers:  
+
Used primers:<br>
−
forward - 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGTATAAAATTTTAATAGT-3' </b>
+
forward - 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGTATAAAATTTTAATAGT-3'<br>
−
reverse - 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTACTTTTTTATTTTTA-3' </b>
+
reverse - 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTACTTTTTTATTTTTA-3'<br>
−
to PCR NisR and NisK from the genome and add the BioBrick prefix and suffix. </b></b>
+
to PCR NisR and NisK from the genome and add the BioBrick prefix and suffix.
  
−
Changed the GTG start codon of NisR into a ATG.
+
 
 +
The GTG start codon of NisR was changed into a ATG, as this is standard with most BioBricks.
 +
 
 +
 
 +
During the making of this BioBrick two mutations occured in the gene. At residue 199 of NisR a GAA was changed to a GAG, not changing the amino acid at this position. At residue 300 of NisK an ACA was changed to an ATA, changing a threonine to an isoleucine.
  
 
===Source===
 
===Source===
  
−
Obtained by PCR on the genome of Lactococcus lactis NZ9700, a nisin producing strain.  
+
Obtained by PCR on the genome of ''Lactococcus lactis'' NZ9700, a nisin producing strain.
  
 
===References===
 
===References===
 +
Trmčić, A. et al. (2011) Complete nisin A gene cluster from ''Lactococcus lactis'' M78 (HM219853) – obtaining the nucleic acid sequence and comparing it to other published nisin sequences. ''Genes Genom.'' 33: 217-221

Latest revision as of 02:41, 18 October 2014


NisR and NisK


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Used primers:
forward - 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGTATAAAATTTTAATAGT-3'
reverse - 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTACTTTTTTATTTTTA-3'
to PCR NisR and NisK from the genome and add the BioBrick prefix and suffix.


The GTG start codon of NisR was changed into a ATG, as this is standard with most BioBricks.


During the making of this BioBrick two mutations occured in the gene. At residue 199 of NisR a GAA was changed to a GAG, not changing the amino acid at this position. At residue 300 of NisK an ACA was changed to an ATA, changing a threonine to an isoleucine.

Source

Obtained by PCR on the genome of Lactococcus lactis NZ9700, a nisin producing strain.

References

Trmčić, A. et al. (2011) Complete nisin A gene cluster from Lactococcus lactis M78 (HM219853) – obtaining the nucleic acid sequence and comparing it to other published nisin sequences. Genes Genom. 33: 217-221